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A strategy for trade monitoring and substitution of the organs of threatened animals Jiao-yang Luo1, Dan Yan1, Jing-yuan Song2, Da Zhang1, Xiao-yan Xing1, Yu-mei Han1, Mei-hua Yang2, Xiao-ping Dong3, Cheng Peng3, Shi-lin Chen2


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A strategy for trade monitoring and substitution of the organs of threatened animals

Jiao-yang Luo1, Dan Yan1, Jing-yuan Song2, Da Zhang1, Xiao-yan Xing1, Yu-mei Han1, Mei-hua Yang2, Xiao-ping Dong3, Cheng Peng3, Shi-lin Chen2, Xiao-he Xiao1
1 China Military Institute of Chinese Medicine, Integrative Medical Center of the 302 Military Hospital, Beijing 100039, P.R. China; 2 Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100094, P.R. China; 3 College of Pharmacy, Chengdu University of Chinese Traditional Medicine, Chengdu 610075, P.R. China

Table S1. GenBank accessions of 241 closely related species used to obtain ADDs; the COI sequences were cropped from the complete coding sequences and analyzed.

GenBank accessions of close-related species of MAHS

AB108504

AB116261

AB245426

AB245427

AF33441

AF547270

AY126697

AY225986

AY239042

AY488491

AY526085

AY676855

AY676856

AY676857

AY676858

AY676859

AY676860

AY676861

AY676862

AY676863

AY676864

AY676865

AY676866

AY676867

AY676868

AY676869

AY676870

AY676871

AY676872

AY676873

AY684273

AY702618

AY858379

DQ124371

DQ124372

DQ124375

DQ124376

DQ124377

DQ124378

DQ124379

DQ124380

DQ124381

DQ124382

DQ124383

DQ124384

DQ124385

DQ124386

DQ124387

DQ124388

DQ124389

DQ124390

DQ124391

DQ124392

DQ124393

DQ124394

DQ124395

DQ124396

DQ124397

DQ124398

DQ124399

DQ124400

DQ124401

DQ124402

DQ124403

DQ124404

DQ124405

DQ124406

DQ124407

DQ124408

DQ124409

DQ124410

DQ124411

DQ124412

DQ124413

DQ124414

DQ124415

DQ124416

DQ124417

DQ124418

DQ347622

DQ347626

DQ989636

EF035447

EF050073

EF494177

EF494178

EF494179

EU177815

EU177816

EU177817

EU177818

EU177819

EU177820

EU177821

EU177822

EU177823

EU177824

EU177825

EU177826

EU177827

EU177828

EU177829

EU177830

EU177831

EU177832

EU177833

EU177834

EU177835

EU177836

EU177837

EU177838

EU177839

EU177840

EU177841

EU177842

EU177843

EU177844

EU177845

EU177846

EU177847

EU177848

EU177849

EU177850

EU177851

EU177852

EU177853

EU177854

EU177855

EU177856

EU177857

EU177858

EU177859

EU177860

EU177861

EU177862

EU177863

EU177864

EU177865

EU177866

EU177867

EU177868

EU177869

EU177870

EU177871

EU315254

EU708434

EU809939

FJ171914

FJ171915

FJ469673

FJ763736

FJ890514

FJ971080

FJ971081

FJ971082

FJ971083

FJ971084

FJ971085

FJ971086

FJ971087

FJ971088

FJ997262

FJ185309

FJ461529

FJ461530

FJ461531

GQ129207

GQ856370

GQ129208

GU068049

GU229278

GU229279

GU229280

GU229281

GU457433

GU457434

GU457435

GU563918

GU985279

HM045017

HM045018

JN190944

JN190945

JN190946

JN315436

JN315437

JN315438

JN315439

JN315440

JN315441

JN315442

JN315443

JN417004

NC_001941

NC_002780

NC_004027

NC_004069

NC_004563

NC_004577

NC_005044

NC_005971

NC_006082

NC_006132

NC_006295

NC_006380

NC_006853

NC_006973

NC_006993

NC_007179

NC_007441

NC_007703

NC_008414

NC_008462

NC_008491

NC_008749

NC_009330

NC_009509

NC_009510

NC_010640

NC_010970

NC_010973

NC_011819

NC_011821

NC_012054

NC_012096

NC_012098

NC_012346

NC_012414

NC_012706

NC_012833

NC_013069

NC_013751

NC_013834

NC_013836

NC_013840

NC_013841

NC_013996

NC_014044

NC_014054

NC_014102

X61145














Table S2. Endotoxin contents in rabbit sera at 1.5 h as determined using a dynamic nephelometric method.




Dilution multiple

Extra endotoxin concentration (EU/mL)

Recovery

(%)


Reaction time (s)

Coefficient of variability (%)

Measured Value (EU/mL)

Reference value (EU/mL)

velvet antler

40

0



3115

0.00

0.0214

0.856

40

0.2

88.15

1258

0.84

0.1947



antler

40

0



>3600

0.00

<0.013

<0.520

40

0.2

100.25

1271

1.53

0.2266



saiga antelope

40

0



>3600

0.00

<0.013

<0.520

40

0.2

93.85

1253

1.24

0.2287



domestic goat

40

0



>3600

0.00

<0.013

<0.520

40

0.2

136.1

1248

3.51

0.2840



negative control

40

0



>3600

0.00

<0.013

<0.520

40

0.2

130.25

1221

2.58

0.2625



Table S3. Damage to spleen lymphocytes (cultured in vitro) under the influence of ADDs.

Concentration

(mg/mL)

The damage extent to spleen lymphocytes (cultured in vitro) influenced by MAHS (%)

velvet antlter

antler

saiga antelope

domestic goat

Asian water buffalo

Chinese pangolin

Chinese softshell turtle

100

35.83

47.38

91.42

59.84

87.95

88.34

85.27

80

0

0

70.86

23.57

47.42

52.37

41.26

60

0

0

38.52

0

21.85

33.47

26.39

40

0

0

0

0

0

0

0

20

0

0

0

0

0

0

0

10

0

0

0

0

0

0

0

5

0

0

0

0

0

0

0

2.5

0

0

0

0

0

0

0

1.25

0

0

0

0

0

0

0

0.625

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Table S4. A review of the status of threatened animals and their substitutes (the substitutes recommended by this study are included).

Scientific name

Common name

IUCN Red List

CITES legislation

Pop. trend

Alternatives and scope

Cetacean

Balaenoptera borealis

Sei whale

Endangered 

Appendix I

Unknown

×

Balaenoptera musculus

Blue whale

Endangered

Appendix I

Increasing

×

Balaenoptera physalus

Fin whale

Endangered

Appendix I

Unknown

×

Physeter macrocephalus

Sperm whale

Vulnerable

Appendix I

Unknown

√ Sperm oil

⊙Jojoba oil

◇Various lubricant applications, wetting agents and extreme pressure additives

⊙Deep water fish species



Hoplostethus atlanticus

Allocyttus sp.

Pseudocyttus maculatus

◇Liquid waxes, used in the cosmetic and high-grade lubricant fields



Balaenoptera bonaerensis

Antarctic minke whale

DD

Appendix I

Unknown

×

Balaenoptera edeni

Bryde's whales

DD

Appendix I

Unknown

×

Balaenoptera omurai

Omura’s whale

DD

Appendix I

Unknown

×

Carnivora
















Panthera tigris

Tiger

Endangered

Appendix I

Decreasing

√Tiger-bone(medical use)

Myospalax baileyi

◇Curerheumatic diseases


Ursus thibetanus

Himalayan black bear

Vulnerable

Appendix I

Decreasing

√Bear bile(medical use)

⊙Pig bile

◇Anti-inflammatory,

anti-convulsion and analgesic effects.

⊙Rabbit bile

◇Positive inotropic action, sedation, anti-tussive action, and anti-histaminic action.

⊙Coptis

△Clear heat, detoxify, purify liver, improve eyesight et al.



Proboscidea

Elephas maximus

Asian elephant

Endangered

Appendix I

Decreasing

√Ivory

⊙Mammoth ivory, cow bone, camel bone

◇Ivory carving


Pholidota

Manis pentadactyla

Chinese pangolin

Endangered

Appendix II

Decreasing

√ pangolin scale(medicinal use)

⊙turtle shell

△Immunoregulatory activity


Perissodactyla

Diceros bicornis

Black rhinoceros

Critically endangered

Appendix I

Increasing

√Rhinoceros horn(medical use)

⊙water buffalo horn

◇Antipyretic effects


Dicerorhinus sumatrensis

Sumatran rhinoceros

Critically endangered

Appendix I

Decreasing

Rhinoceros sondaicus

Javan rhinoceros

Critically endangered

Appendix I

Unknown

Rhinoceros unicornis

Indian rhinoceros

Vulnerable

Appendix I

Increasing

Ceratotherium simum

White rhinoceros

Near threatened

Appendix I,II

Increasing

Artiodactyla

Elaphurus davidianus

Père David's deer

Extinct in the wild

NL

Increasing

×

Bos sauveli

Kouprey

Critically endangered

Appendix I

Unknown

×


Bubalus mindorensis

Tamaraw

Critically endangered

Appendix I

Decreasing

×

Nanger dama

Dama gazelle

Critically endangered

Appendix I

Decreasing

×

Saiga tatarica

Mongolian saiga

Critically endangered

Appendix II

Decreasing

√Antelope's horn( medical use)

⊙goat horn

◇Immunological activity, sedation, analgesia

△ Antipyretic activity



Pseudoryx nghetinhensis

Saola

Critically endangered

Appendix I

Decreasing

×

Axis porcinus

Hog deer

Endangered

Appendix I

Decreasing

×

Axis calamianensis

Calamian hog deer

Endangered

Appendix I

Decreasing

×

Gazella leptoceros

Slender-horned gazelle

Endangered

Appendix I

Decreasing

×

Bubalus depressicornis

Lowland anoa

Endangered

Appendix I

Decreasing

×

Hippocamelus bisulcus

Chilean guemal

Endangered

NL

Decreasing

×

Bosjavanicus

Banteng

Endangered

NL

Decreasing

×

Pantholops hodgsonii

Chiru

Endangered

Appendix I

Decreasing

×

Capra falconeri

Markhor

Endangered

Appendix I

Decreasing

×

Bubalus arnee

Indian water buffalo

Endangered

Appendix III

Decreasing

×

Moschus berezovskii

Forest musk deer

Endangered

Appendix I,II

Decreasing

√Musk(medical use)

Moschus chrysogaster

Alpine musk deer

Endangered

Appendix I,II

Decreasing

Ondatra zibethica

◇Anti-inflammation and analgesion97



Moschus moschiferus

Siberian musk deer

Vulnerable

AppendixI,II

Decreasing

Bos mutus

Wild yak

Vulnerable

Appendix I

Decreasing

√Yak horn(medical use)

Bubalus bubalis

△Anticogulate activity


Bison bonasus

European bison

Vulnerable

NL

Increasing

×

Oryx leucoryx

Arabian oryx

Vulnerable

Appendix I

Stable

×

Eudorcas thomsonii

Thomson's gazelles

Near threatened

NL

Decreasing

×

Procapra gutturosa

Mongolian gazelle

Least concern

NL

Unknown

×

Dama dama

Fallow deer

Least concern

Appendix I

Unknown

×

Cervus nippon

Sika deer

Least concern

NL

Increasing

Pilos deer horn(medical use)

⊙Calcinated antler cancellous bone



◇Source of bone graft material

Cervus elaphus hanglu

Red deer

Least concern

Appendix I

Increasing

×

Cervus elaphus bactrianus

Red deer

Least concern

Appendix II

Increasing

×

Cervus elaphus barbarous

Red deer

Least concern

Appendix III

Increasing

×

Reptile

Eretmochelys imbricata

Hawksbill turtle

Critically endangered

NL

Decreasing

×

Pelochelys cantorii

Frog-faced softshell turtle

Endangered

Appendix II

Needs updating

×

Platysternon megacephalum

Big-headed turtle

Endangered

Appendix II

Needs updating

×

Caretta caretta

Loggerhead

Endangered

NL

Needs updating

×

Shark

Sphyrna lewini

Scalloped hammerhead shark

Endangered

Appendix III

Unknown

×

Cetorhinus maximus

Basking shark

Vulnerable

Appendix II

Decreasing

×

Carcharodon carcharias

Great white shark

Vulnerable

Appendix II

Unknown

×

DD: data deficient; NL: not listed; √ medicinal parts or other applications; × no alternative use; ⊙substitutes; ◇alternative application scope; △ penitential recommendation by the present study.


Figure S1. Distribution of some endangered and threatened species examined in this study. (A) General distribution of the species in the wild; (B) detailed distribution of endangered and threatened species. (The information of the maps was derived from investments made in TM in China, and we hold the copyright of this figure.)



Figure S2. Neighbor-joining tree based on Kimura 2-parameter distances for 46 COI gene sequences.



Figure S3. Barcoding gap test. The relative distribution of inter- and intraspecific variations.



Figure S4. Antipyretic activities of ADDs. (A) Maximum changes in body temperature in the presence of various ADDs; (B) PGE2 level in rabbit serum; (C) PGE1 level in rabbit serum; (D) TNF-α level in rabbit serum; (E) d-values among the temperatures.



Figure S5. The effects of ADDs on liver microsomes. (A) HFP-t curves of the growth of liver microsomes in the presence of ADDs in vitro, within which (a)-(e) represent velvet antler group, antler group, blank control, domestic group and saiga antelope group, respectively; (B) total heat output of the heat-flow power under the action of various ADDs; (C) inhibition ratios of the ADDs on the growth of liver microsomes in vitro.



Figure S6. The effects of various ADDs on spleen lymphocytes. (A) Kunming mice; (B) cultured spleen lymphocytes; (C) microcalorimetry; (D) HFP-t curves of the growth and metabolism of spleen lymphocytes in the presence of ADDs, within which (a)-(h) represent velvet antler group, domestic goat group, antler group, blank group, Chinese pangolin group, saiga antelope group, Asian water buffalo group, and Chinese softshell turtle group, respectively; (E) parameters extracted from the HFP-t curves; (F) comparison of the values of the heat-flow power in the presence of various ADDs; (G) comparison of the values of the total heat output in the presence of various ADDs.



Figure S7. Investigation of the temperature gradients in the mouse thermotaxis study.



Figure S8. Investigation of the observation time in the mouse thermotaxis study.


Figure S9. The effects of ADDs on mouse thermotaxis. (A) Temperature-controlled setup; (B) remote monitoring system; (C) data-processing system; (D): comparison of the proportion of animals in the warm zone in the presence of various ADDs; (E) comparison of the 30 min moving distances in the presence of various ADDs; (F) comparison of times taken to cross from one zone to another in the presence of various ADDs; (G) 5-HT levels in mouse sera; (H) cAMP levels in mouse sera; (I) cGMP levels in mouse sera; (J) cAMP/cGMP values.


Figure S10. Determination of 5-HT, cAMP, and cGMP levels in the brain tissues of mice. (A) 5-HT level in the brain tissues of mice; (B) cAMP level in the brain tissues of mice; (C) cGMP level in the brain tissues of mice; (D) cAMP /cGMP values.



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